高效液相色谱-串联质谱法测定食品原料中烟酰胺单核苷酸的含量

Determination of Nicotinamide Mononucleotide in the Natural Food Materials by High Performance Liquid Chromatography-Mass Spectrometry

建立烟酰胺单核苷酸的高效液相色谱-串联质谱测定方法,分析不同食品原料中烟酰胺单核苷酸的含量。样品经5%甲醇水溶液提取,采用Shim-pack WAX-2色谱柱分离,流动相为乙腈与5%乙腈水溶液(含20 mmol/L乙酸铵、20 mmol/L氨水),梯度洗脱15 min,流速为0.5 mL/min,柱温为30 ℃,在质谱正离子模式下采用多反应监测模式进行测定,定量离子对为m/z335.0/123.0。在该条件下,烟酰胺单核苷酸在10~1000 μg/L质量浓度范围内线性良好(R^(2)=0.9999),检出限为5.0 μg/L,定量限为10.0 μg/L,回收率为90.14%~105.49%,相对标准偏差为1.47%~4.82%。该方法前处理简单、选择性高、回收率良好,适用于不同种类食品原料中烟酰胺单核苷酸的定量分析。采用该方法分析发现,不同食品原料中烟酰胺单核苷酸含量存在明显差异,番茄、香菇、黄豆、鳄梨、对虾、南极磷虾是其良好的天然食物来源。研究结果可为烟酰胺单核苷酸相关膳食营养搭配、产品开发与质量评价提供理论指导和技术支撑。

A rapid determination method of nicotinamide mononucleotide by high performance liquid chromatography tandem mass spectrometry was established and used to analyze the NMN contents of various natural food materials in the present study.The samples were extracted by 5% methanol aqueous solution and then separated by Shim-pack WAX-2 column at 30 ℃.The mobile phase was composed of the gradient changes of acetonitrile and 5% acetonitrile aqueous solution (containing 20 mmol/L ammonium acetate and 20 mmol/L ammonia) for 15 min at a flow rate of 0.5 mL/min.Quantitative determination was performed at the multi reaction monitoring mode under the positive ion mode of mass spectrometry and the quantitative ion pair was m/z 335.0/123.0.The method showed a good linear relationship between peak area and concentration over the range from 10 μg/L to 1000 μg/L (R^(2)=0.9999).The limit of detection was 5.0 μg/L and the limit of quantitation was 10.0 μg/L.The recoveries of nicotinamide mononucleotide at three concentration levels ranged from 90.14%~105.49% and the relative standard deviation of precision was 1.47%~4.82%.This method possessed the advantages of simple pretreatment,high selectivity and good recovery,which was suitable for the quantitative analysis of nicotinamide mononucleotides in different kinds of food materials.There were marked differences in the nicotinamide mononucleotide contents of the different food materials.Tomato,mushroom,soybean,avocado,prawn and antarctic krill were the good natural food sources of nicotinamide mononucleotide.This research could provide theoretical guidance and technical support for nicotinamide mononucleotide related dietary nutrition collocation,product development and quality evaluation.